Motor behaviors such as locomotion rely on precise signaling from the nervous system to coordinate various muscle activities. The neuromuscular junction has been extensively studied and a considerable amount of information exists on how information is communicated across the synapse between a motor neuron and a muscle cell. At the cellular level, depolarization of the motor neuron produces an action potential that is propagated to the presynaptic terminal. Depolarization causes the opening of voltage-gated calcium channels at the presynaptic terminal, resulting in an increase in intracellular calcium. In response to local increases in calcium, neurotransmitter filled vesicles fuse with the presynaptic plasma membrane, releasing their contents into the synaptic cleft. Postsynaptic ligand-gated ion channels on the muscle bind the neurotransmitter, resulting in the opening of an integral ion channel. Depending on an ion channel's permeability, the muscle is either hyperpolarized or depolarized.
Chemical synaptic transmission mediates fast and slow communication between cells, but it is not the only means of communication within the nervous system. Signals may also be conveyed nonsynaptically. In this case, a neurotransmitter or hormone is released at one site and slowly diffuses to distant target sites that are not in contact with the original site of release. This type of signaling is more suited to global, modulatory activities rather than functions requiring a rapid response. The interplay of synaptic and non-synaptic communication ultimately determines how a nervous system mediates particular behaviors.
In C. elegans, synaptic transmission has been extensively studied at the genetic, cellular and molecular levels. Most behaviors in the worm follow a simple paradigm in which information is directed from the sensory neurons to motor neurons to muscle, via synaptic contacts. However, the defecation cycle in C. elegans is a unique behavior in that it appears to be mediated by both synaptic, as well as nonsynaptic transmission (McIntire et al., 1993b; Thomas, 1990). Defecation in C. elegans is a stereotyped behavior that occurs every 50 seconds for the life of the animal, and is characterized by the coordinated activation of three independent muscle contractions (Croll, 1975; Thomas, 1990). The cycle is initiated with a posterior body contraction, followed by an anterior body contraction and finalized by an enteric muscle contraction, which expels intestinal contents. A simplified genetic pathway to explain the defecation behavior was proposed by Jim Thomas (Thomas, 1990) (see, FIG. 1B). In this model, a clock mechanism keeps time independently of the motor program. At the appropriate interval, the clock first signals the posterior body contraction and then signals the common anterior body and enteric muscle contraction mechanism, which ultimately leads to activation of the individual muscle contractions (Liu and Thomas, 1994; Thomas, 1990) (FIG. 1B).
To determine the cellular basis of the defecation motor program, extensive cellular laser ablations have been performed. Based on these studies the motor neurons AVL and DVB were demonstrated to mediate the anterior body and enteric muscle contraction, but not posterior body contraction (McIntire et al., 1993b). The anterior body contraction is mediated by the motor neuron AVL, but the neurotransmitter that mediates this contraction is unknown (see, FIG. 1A). While AVL alone is required for anterior body contraction, both the AVL and DVB motor neurons serve a redundant function in activating the enteric muscles (see, FIG. 1A). Activation of the enteric muscles is GABA-dependent and is mediated by the EXP-I receptor (Beg and Jorgensen, 2003; McIntire et al., 1993a; McIntire et al, 1993b).
Interestingly, no known neurons are required to maintain the clock or initiate the posterior body contraction, suggesting that cycle timing and posterior body contraction are mediated by a non-neuronal mechanism (Liu and Thomas, 1994; McIntire et al., 1993b; Thomas, 1990) (see, FIG. 1A). Furthermore, mutations that disrupt classical neurotransmission and secretion do not affect the posterior body contraction. Taken together, these data suggest that posterior body contraction occurs through a non-neuronal mechanism that does not rely on classical or peptidergic neurotransmission.
Many genes have been identified that affect only specific aspects of the defecation motor program. It has been demonstrated that timekeeping of the cycle is controlled by an endogenous clock that resides in the intestine (Dal Santo et al., 1999). The cycle time is set by the activity of the itr-1 gene, which encodes an inositol triphosphate (IP3) receptor which mediates release of calcium from the smooth endoplasmic reticulum into the intestine every 50 seconds (Dal Santo et al., 1999). Mutations in the itr-1 gene slow down or eliminate the cycle, while overexpression accelerates the cycle. In the intestine, calcium levels oscillate with the same period as the defecation cycle (50 seconds) and peak calcium levels immediately precede the posterior body contraction (Dal Santo et al., 1999). Therefore, the frequency of intracellular calcium release in the intestine, determines the frequency of the defecation cycle.
It has been demonstrated that there is a one-to-one relationship between the calcium spike in the intestine and the execution of the posterior body contraction (Dal Santo et al., 1999). Normally, motor behaviors such as muscle contraction are mediated by the nervous system. Since neuronal input is not required to initiate the posterior body contraction, these muscles could be directly activated by a Ca2+ regulated signal from the intestine.